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Protein Phosphatases Assay Kits

iD Sensolyte® Assay Kits

Alkaline phosphatases catalyze the dephosphorylation of biomolecules in an alkaline environment and play a key role in cell signal transduction. The change of alkaline phosphatase activity is involved in a variety of physiological and pathological events, such as bone development, bone-related diseases, gestation-related diseases and inflammatory bowel diseases. Alkaline phosphatase is also a popular enzyme conjugated with secondary antibody for ELISA and serves as a reporter for gene expression.

p-Nitrophenyl phosphate (pNPP) is proven to be an effective chromogenic substrate for alkaline phosphates.

FDP (3,6-fluorescein diphosphate) is a highly sensitive fluorogenic substrate for most phosphatases, i.e., alkaline phosphatases, protein tyrosine phosphatases and serine/threonine phosphatases. The final hydrolytic product of FDP is fluorescein, a fluorophore that has a very high emission quantum yield. The signal can easily be read by a fluorescence plate reader at Ex/Em=485±20/528±20 nm. The fluorescence of FDP hydrolyzed product is strengthened when pH > 7.0. For some protein phosphatase requiring slightly acidic buffer for reaction, a pH-adjusting stop solution should be added before reading the fluorescence signal.

MFP is a proprietary fluorogenic substrate for most phosphatases, i.e., alkaline phosphatases, acid phosphatases, protein tyrosine phosphatases, and serine/threonine phosphatases. The phosphatase-induced hydrolysis of MFP yields an intense fluorescent product that has excitation wavelength around 488 nm, and maximum emission around 520 nm. This characteristic makes the substrate adaptable to all the fluorescence instruments. The fluorescence of hydrolyzed product of MFP is pH-insensitive, thus it can be used to continuously monitor phosphatase activity in acidic buffer.

The secreted alkaline phosphatase (SEAP) is widely used as reporter gene to analyze the activity of promoters and transcriptional factors and to tract gene expression in cell culture or animals. SEAP can be effectively secreted into culture medium and serum. This unique characteristic of SEAP makes it convenient to kinetically analyze gene expression over a period of time using a single culture or animal.

MG (Malachite Green) is useful to monitor phosphate moity, based on the quantification of the green complex formed between Malachite Green, molybdate, and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600-660 nm) or on a plate reader.

DescriptionSize Reference USD Qty  
SensoLyte® FDP Alkaline Phosphatase Assay Kit *Fluorimetric* - 1 kit1 kit AS-71109 286.00Add to cart
SensoLyte® FDP Alkaline Phosphatase ELISA Assay Kit *Fluorimetric* - 1 kit1 kit AS-71101-M 286.00Add to cart
SensoLyte® FDP Alkaline Phosphatase ELISA Assay Kit *Fluorimetric* - 1 kit1 kit AS-71101-R 286.00Add to cart
SensoLyte® FDP Protein Phosphatase Assay Kit *Fluorimetric* - 1 kit1 kit AS-71100 286.00Add to cart
SensoLyte® Luminescent Alkaline Phosphatase Assay Kit *Luminometric* - 1 kit1 kit AS-72122 356.00Add to cart
SensoLyte® Luminescent Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Luminometric* - 1 kit1 kit AS-72145 356.00Add to cart
SensoLyte® MFP Acid Phosphatase Assay Kit *Fluorimetric* - 1 kit1 kit AS-71231 310.00Add to cart
SensoLyte® MFP Protein Phosphatase Assay Kit *Fluorimetric* - 1 kit1 kit AS-71104 286.00Add to cart
SensoLyte® MG Phosphate Assay Kit *Colorimetric* - 1 kit1 kit AS-71103 286.00Add to cart
SensoLyte® pNPP Alkaline Phosphatase Assay Kit *Colorimetric* - 1 kit1 kit AS-72146 182.00Add to cart
SensoLyte® pNPP Protein Phosphatase Assay Kit *Colorimetric* - 1 kit1 kit AS-71105 255.00Add to cart
SensoLyte® pNPP Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Colorimetric* - 1 kit1 kit AS-72144 182.00Add to cart

The SensoLyte® FDP Alkaline Phosphatase ELISA Assay Kits *Fluorimetric* (Rabbit, Mouse) use highly purified FDP to quantify alkaline phosphatase activity in ELISA. They allow to detect as low as atto molar (10-15 M) concentration of test antibody in an ELISA format. The kits contain: • FDP phosphatase substrate (Ex/Em=485±20/528±20 nm upon dephosphorylation) • Assay buffer • Stop solution • Wash buffer • Alkaline phosphatase-conjugated secondary antibody (goat anti-rabbit IgG or goat-anti-mouse IgG) • An optimized ELISA protocol (1 kit = 500 assays).

The SensoLyte® Luminescent Alkaline Phosphatase ELISA Assay Kit *Luminometric* provides a highly sensitive chemiluminescent substrate for quantifying alkaline phosphatase activity in ELISA. The kit contains: • Chemiluminescent alkaline phosphatase substrate • Assay buffer • Wash buffer • Stop solution • An optimized ELISA protocol (1 kit = 500 assays).

The SensoLyte® pNPP Alkaline Phosphatase ELISA Assay Kits *Colorimetric* (Rabbit, Mouse, Goat) provide a sensitive chromogenic substrate for quantifying alkaline phosphatase activity in ELISA. The kits contain: pNPP chromogenic liquid substrate (absorption maximum = 405 nm upon dephosphorylation), Assay buffer, ELISA wash buffer, Stop solution, Alkaline phosphatase conjugated secondary antibody (goat anti-rabbit IgG or goat-anti-mouse IgG or rabbit-anti-goat IgG), an optimized ELISA assay protocol (1 kit = 500 assays).

The SensoLyte® pNPP Alkaline Phosphatase Assay Kit *Colorimetric* can be used to detect alkaline phosphatase activity in biological samples and alkaline phosphatase-based ELISA. The kit contains: pNPP chromogenic liquid substrate (absorption maximum = 405 nm upon dephosphorylation), Assay buffer, Stop solution, Alkaline phosphatase standard, “mix and read” assay protocol that is compatible with high throughput screening (1 kit = 500 assays).

The SensoLyte® pNPP Protein Phosphatase Assay Kit *Colorimetric* can be used for characterizing kinetics of enzyme reaction and high throughput screening of protein phosphatase inhibitors. It has high sensitivity and wide linear range (The detection limit is generally 3 ng or below). The kit contains: • pNNP chromogenic substrate • Protein phosphatase inhibitor • Assay buffer • A 'mix and read' assay protocol that is compatible with HTS liquid handling instruments (1 kit = 500 assays).

The SensoLyte® FDP Alkaline Phosphatase Assay Kit *Fluorimetric* uses highly purified FDP to quantify alkaline phosphatase activity in solutions, in cell extracts, in live cells as well as on solid surfaces (such as PVDF membranes). The kit contains: FDP phosphatase substrate (Ex/Em=485±20/528±20 nm upon dephosphorylation), Calibration standard, Reaction buffer, Stop buffer, An optimized ‘mix and read’ assay protocol that is compatible with HTS liquid handling instruments (1 kit = 500 assays).

The SensoLyte® FDP Protein Phosphatase Assay Kit *Fluorimetric* can be used for characterizing kinetics of enzyme reaction and high throughput screening of protein phosphatase inducers and inhibitors (1 kit = 500 assays).

The SensoLyte® Luminescent Alkaline Phosphatase Assay Kit *Luminometric* provides highly sensitive chemiluminescent substrate to quantify alkaline phosphatase activity in solutions, in cell extracts, in live cells. The kit contains: chemiluminescent substrate, positive control (calf intestine alkaline phosphatase), buffer, stop solution, an optimized ‘mix and read’ assay protocol that is compatible with HTS liquid handling instruments (1 kit = 500 assays).

The SensoLyte® MFP Acid Phosphatase Assay Kit *Fluorimetric* uses MFP to quantify acid phosphatase activities. The assay is highly sensitive and has large dynamic range. The kit contains: MFP fluorogenic substrate (Ex/Em=488 ±20 nm/520 ± 20 nm upon dephosphorylation), Assay buffer, A “mix and read” assay protocol that is compatible with high throughput screening liquid handling instruments (1 kit = 500 assays).

The SensoLyte® MFP Protein Phosphatase Assay Kit *Fluorimetric* uses MFP to quantify protein phosphatase activities. The kit can be used for characterizing kinetics of enzyme reaction and high throughput screening of protein phosphatase inhibitors. The kit contains: • MFP phosphatase substrate • Protein phosphatase inhibitor • Assay buffer • A 'mix and read' assay protocol that is compatible with HTS liquid handling instruments (1 kit = 500 assays).

The SensoLyte® MG Phosphate Assay Kit *Colorimetric* is based on quantification of the green complex formed between MG (Malachite Green), molybdate, and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600-660 nm) or on a plate reader. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be performed in tubes, cuvettes, or multi-well plates. The assays can be conveniently performed in 96 or 384-well plates for high-throughput screening of enzyme inhibitors. The kit has been used for assaying phosphatase, phospholipase or lipase, nucleoside triphosphatase (ATPase, GTPase, etc.) and phytase. The kit has the following components: • Highly purified Malachite Green reagent • Protein phosphatase inhibitor • Assay buffer • A 'mix and read' assay protocol that is compatible with HTS liquid handling instruments with Z' factors of 0.7 to 0.9 (1 kit = 1000 assays).

The SensoLyte® FDP Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Fluorimetric* provides a sensitive fluorimetric assay of placental alkaline phosphatase for both secreted and membrane-bound form of placental alkaline phosphatases. The kit contains: FDP fluorogenic substrate (Ex/Em=485±20/528±20 nm upon dephosphorylation), Assay buffer, Stop solution, A “mix and read” assay protocol that is compatible with high throughput screening liquid handling instruments (1 kit = 500 assays).

The SensoLyte® Luminescent Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Luminometric* provides a highly sensitive assay of placental alkaline phosphatase for both secreted and membrane-bound form of enzyme. The kit contains: Chemiluminescent substrate, Assay buffer, Stop solution, Placental alkaline phosphatase standard, “mix and read” assay protocol that is compatible with high throughput screening (1 kit = 500 assays).

The SensoLyte® pNPP Secreted Alkaline Phosphatase Reporter Gene Assay Kit *Colorimetric* provides a sensitive colorimetric assay of placental alkaline phosphatase for both secreted and membrane-bound form of enzyme. The kit contains: pNPP chromogenic liquid substrate (absorption maximum = 405 nm upon dephosphorylation), Assay buffer, Stop solution, Placental alkaline phosphatase standard, “mix and read” assay protocol that is compatible with high throughput screening (1 kit = 500 assays).
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